KR20070093427A - Stilbene derivatives and their use for binding and imaging amyloid plaques - Google Patents

Abstract

This invention relates to a method of imaging amyloid deposits and to labeled compounds, and methods of making labeled compounds useful in imaging amyloid deposits. This invention also relates to compounds, and methods of making compounds for inhibiting the aggregation of amyloid proteins to form amyloid deposits, and a method of delivering a therapeutic agent to amyloid deposits.

Description

STILBENE DERIVATIVES AND THEIR USE FOR BINDING AND IMAGING AMYLOID PLAQUES}

The present invention relates to novel biologically active compounds, diagnostic imaging methods using radiolabeled compounds and methods of preparing radiolabeled compounds.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive decline, irreversible memory loss, disorientation and impaired speech. Post-mortem examination of AD brain sections shows abundant senile plaques (SPs) consisting of amyloid-β (Aβ) peptides and numerous neurofibrillary tangles (NFTs) formed by high levels of phosphorylated tau protein fibers (recently For review articles and additional citations, see Ginsberg, SD, et al., "Molecular Pathology of Alzheimer's Disease and Related Disorders," in Cerebral Cortex: Neurodegenerative and Age-related Changes in Structure and Function of Cerebral Cortex, Kluwer Academic / Plenum, NY (1999), pp. 603-654; Vogelsberg-Ragaglia, V., et al., "Cell Biology of Tau and Cytoskeletal Pathology in Alzheimer's Disease," Alzheimer's Disease, Lippincot, Williams & Wilkins, Philadelphia, PA ( 1999, pp. 359-372).

Amyloidosis is a disease characterized by the deposition of various insoluble fibrous proteins in the tissues of a patient. Amyloid deposits are formed by aggregation of amyloid proteins and then further combinations of aggregates and / or amyloid proteins. Formation and deposition of β-amyloid (Aβ) peptide aggregates in the brain are important factors for the development and progression of AD.

In addition to the role of amyloid deposits in Alzheimer's disease, the presence of amyloid deposits can be attributed to Mediterranean fever, Muckle-Wells syndrome, idiopathic myeloma, amyloid polyneuropathy, amyloid cardiomyopathy, systemic senile amyloidosis, amyloid polyneuropathy, Amyloid Hereditary Cerebral Hemorrhage, Down's Syndrome, Scrapie, Creutzfeldt-Jakob Disease, Kurou Disease, Gerstmann-Straussler-Schainker Syndrome, Thyroid Medulla, Isolated Arterial Amyloid, β ₂ in Dialysis Patients- It has been shown in diseases such as microglobulin amyloid, inclusion body myositis, β ₂ -amyloid deposits in myasthenia, and Langerhans Island type II diabetic insulin species.

Fibrous aggregates of amyloid peptides Aβ _1-40 and Aβ _1-42 are major metabolic peptides derived from amyloid precursor proteins found in senile plaques and cerebrovascular amyloid deposits in AD patients (Xia, W., et al., J. Proc. Natl. Acad. ScL USA 97: 9299-9304 (2000)). Prevention and recovery of Αβ plaque formation has been targeted as a treatment for such diseases (Selkoe, D., J. JAMA 283: 1615-1617 (2000); Wolfe, MS, et al., J. Med. Chem. 41: 6-9 (1998); Skovronsky, DM, and Lee, VM, Trends Pharmacol.Sci 21: 161-163 (2000)).

Familial AD (FAD) is caused by a number of mutations in the A precursor protein (APP), presenilin 1 (PS1) and presenilin 2 (PS2) genes (Ginsberg, SD, et al., “Molecular Pathology of Alzheimer's Disease and Related Disorders, "in Cerebral Cortex: Neurodegenerative and Age-related Changes in Structure and Function of Cerebral Cortex, Kluwer Academic / Plenum, NY (1999), pp. 603-654; Vogelsberg-Ragaglia, V., et al., "Cell Biology of Tau and Cytoskeletal Pathology in Alzheimer's Disease," Alzheimer's Disease, Lippincot, Williams & Wilkins, Philadelphia, PA (1999), pp. 359-372).

Although the exact mechanisms inherent in AD are not fully known, all pathogenic FAD mutations studied to date have increased production of Αβ peptides with extended amyloid-producing amino acids 42-43. Thus, at least in FAD, deregulation of Αβ production appears to be sufficient to induce a cascade of a series of events leading to neurodegeneration. Indeed, the amyloid cascade hypothesis suggested that the formation of extracellular fibrous Aβ aggregates in the brain may be a central event in AD pathogenesis (Selkoe, DJ, "Biology of β-amyloid Precursor Protein and the Mechanism of Alzheimer's Disease, '' Alzheimer's Disease, Lippincot Williams & Wilkins, Philadelphia, PA (1999), pp. 293-310; Selkoe, DJ, J. Am. Med. Assoc. 283: 1615-1617 (2000); Naslund, J., et al J. Am. Med. Assoc. 283: 1571-1577 (2000); Golde, TE, et al., Biochimica et Biophysica Acta 1502: 172-187 (2000).

Various approaches to inhibit the production of fibrous Αβ and reduce deposition in the brain are currently being evaluated as potential therapies for AD (Skovronsky, DM and Lee, VM, Trends Pharmacol. Sci 17: 161-163 (2000) Vassar, R., et al., Science 286: 735-741 (1999); Wolfe, MS, et al., J. Med. Chem. 41: 6-9 (1998); Moore, CL, et al. 43: 3434-3442 (2000); Findeis, MA, Biochimica et Biophysica Acta 1502: 76-84 (2000); Kuner, P., Bohrmann, et al., J. Biol. Chem. 275: 1673-1678 (2000). Therefore, there is interest in developing ligands that specifically bind to fibrous Αβ aggregates. Since extracellular SP is an accessible target, these novel ligands could be used as diagnostic tools in vivo and as probes for the visualization of progressive deposition of Aβ in AD amyloidogenesis studies of surviving patients.

For this purpose, several interesting approaches have been reported for developing fibrous Aβ aggregate-specific ligands (Ashburn, TT, et al., Chem. Biol 3: 351-358 (1996); Han, G., et al. J. Am. Chem. Soc. 118: 4506-4507 (1996); Klunk, WE, et al., Biol. Psychiatry 35: 627 (1994); Klunk, WE, et al., Neurobiol. 541-548 (1995); Klunk, WE, et al., Society for Neuroscience Abstract 23: 1638 (1997); Mathis, CA, et al., Proc. XIIth Intl. Symp. Radiopharm. Chem., Uppsala, Sweden: 94-95 (1997); Lorenzo, A. and Yankner, BA, Proc. Natl Acad. Sci. USA 91: 12243-12247 (1994); Zhen, W., et al., J. Med. Chem. 42: 2805-2815 (1999). The most attractive approach is based on highly conjugated chrysamine-G (CG) and Congo Red (CR), the latter being used for fluorescence staining of SP and NET in post AD brain sections (Ashburn, TT, et al., Chem. Biol. 3: 351-358 (1996); Klunk, WE, et al., J. Histochem. Cytochem. 37: 1273-1281 (1989)). Inhibition constants (K _i ) of binding to fibrous Aβ aggregates of 3'-bromo- and 3'-iodo derivatives of CR, CG and CG are 2,800, 370, 300 and 250 nM, respectively (Mathis, CA, et al., Proc. XIIth Intl. Symp. Radiopharm. Chem., Uppsala, Sweden: 94-95 (1997). These compounds have been shown to selectively bind not only to fibrous Aβ deposits of AD brain sections but also to in vitro Aβ (1-40) peptide aggregates (Mathis, CA, et al., Proc. XIIth Intl. Symp. Radiopharm. Chem., Uppsala, Sweden: 94-95 (1997).

Imaging Aβ aggregates in the brain has several potential advantages. Imaging techniques will improve diagnosis by identifying potential patients with excess Aβ plaques in the brain, and therefore, these patients may be more likely to develop Alzheimer's disease. The technique will also be useful for monitoring disease progression. Once anti-plaque drug treatment is available, imaging of Αβ plaques in the brain can provide an important tool for monitoring treatment. Thus, there is an urgent need for simple non-invasive methods for detecting and quantifying amyloid deposits in patients. Currently, detection of amyloid deposits includes histological analysis of biopsy or autopsy. Both methods have disadvantages. For example, an autopsy can only be used for post diagnosis.

It is difficult to image amyloid deposits directly in vivo because the deposits have many of the same physical properties as normal tissues (eg density and moisture content). Attempts to image amyloid deposits using magnetic resonance imaging (MRI) and computed tomography (CAT) have been disappointing and amyloid deposits were detected only under certain favorable conditions. In addition, efforts to label amyloid deposits with antibodies, serum amyloid P proteins, or other probe molecules provided some selectivity for tissue periphery, but poor imaging for tissue interiors.

Potential ligands for detecting Aβ aggregates in the surviving brain must cross the intact blood-brain barrier. Thus, brain uptake can be improved by using ligands having relatively small molecular size (compared to Congo Red) and high lipophilicity. Highly conjugated thioflavin (S and T) is commonly used as a dye for staining Aβ aggregates in the AD brain (Elhaddaoui, A., et al., Biospectroscopy 1: 351-356 (1995)).

[ ¹⁸ F] FDDNP, a high-lipophilic tracer for binding to both knots (mainly composed of overphosphorylated tau protein) and plaques (containing Aβ protein aggregates), has been reported (Shoghi-Jadid K, et al., Am J Geriatr Psychiatry. 2002; 10: 24-35). Using positive electron emission tomography (PET), it has been reported that the tracer specifically labels plaques and knot deposits in nine AD patients and seven comparison subjects (Nordberg A. Lancet Neurol. 2004 ; 3: 519-27). A new pharmacokinetic assay called the relative residence time of the subject brain area versus the pons was used to illustrate the difference between AD patients and the comparison subjects. Relative residence time was significantly higher in AD patients. This is further complicated by the interesting finding that FDDNP competes with several NSAIDs for binding to Aβ fibers in vitro and to Aβ plaques in vitro (Agdeppa ED, et al. 2001; Agdeppa ED, et al., Neuroscience. 2003). 117: 723-30).

It has recently been reported to image β-amyloid in the brain of AD patients using the benzothiazole aniline derivative [ ¹¹ C] 6-OH-BTA-1 (also referred to as [ ¹¹ C] PIB) (Mathis CA, et al. , Curr Pharm Des. 2004; 10: 1469-92; Mathis CA, et al., Arch. Neurol. 2005, 62: 196-200.). In contrast to that observed for [ ¹⁸ F] FDDNP, [ ¹¹ C] 6-OH-BTA-1 selectively binds to fibrous Αβ in vivo. Patients diagnosed with mild AD showed marked retention of [ ¹¹ C] 6-OH-BTA-1 in the cortex known to contain large amounts of amyloid deposits in AD. In the AD patient group, [ ¹¹ C] 6-OH-BTA-1 retention was most significantly increased in the frontal cortex. In addition, significant increases were observed in the parietal, temporal and occipital cortex and striatum. [ ¹¹ C] 6-OH-BTA-1 retention was equivalent in comparable subjects to AD patients in areas known to be relatively unaffected by amyloid deposition (eg, subcortical white matter, glial and cerebellum). Recently, another ¹¹ C labeled Aβ plaque-targeting probe, Stilbene derivative- [ ¹¹ C] SB-13, has been studied. In vitro binding with [ ³ H] SB-13 suggested that the compound exhibits good binding affinity and that binding can be clearly measured in cortical gray matter but not in the white matter of AD cases (Kung MP, et. al., Brain Res. 2004; 1025: 98-105.). Cortical tissue homogenates of control brains showed very low specific binding. The Kd value of [ ³ H] SB-13 in AD cortical homogenate was 2.4 ± 0.2 nM. High binding capacity and corresponding values were observed (14-45 pmol / mg protein) (same literature). As expected, [ ¹¹ C] SB-13 in AD patients exhibited much deposition in the frontal cortex (expected to contain areas of high density Aβ plaques) in mild to moderate AD patients, but with age-matched Not in the control subjects (Verhoeff NP, et al., Am J Geriatr Psychiatry. 2004; 12: 584-95).

It would be useful to have a non-invasive technique for imaging and quantifying amyloid deposits in a patient. It would also be useful to have a method that demonstrates the ability of a compound to inhibit amyloid protein aggregation to form amyloid deposits and a compound that inhibits amyloid protein aggregation.

Summary of the Invention

The present invention provides novel compounds of formulas (I), (II) and (II).

The present invention also provides a diagnostic composition comprising a radiolabeled compound of formula (I), (II) and (II) and a pharmaceutically acceptable carrier or diluent.

We also provide a method of imaging amyloid deposits, comprising introducing into a patient a detectable amount of a labeled compound of Formula (I), (II) or (III), a pharmaceutically acceptable salt, ester, amide or drug precursor thereof. do.

The present invention also provides a method of inhibiting aggregation of amyloid protein comprising administering to a mammal an amyloid inhibitory amount of a compound of formula (I), (II) or (III), a pharmaceutically acceptable salt, ester, amide, or drug precursor thereof. to provide.

A further aspect of the invention relates to methods of synthesizing amyloid inhibitory and imaging compounds of formula (I), (II) or (III) described herein and intermediates useful therein.

1 shows Ki binding data of some compounds of the invention.

In a first aspect, the invention relates to a compound of formula (I), a pharmaceutically acceptable salt or drug precursor thereof.

Where

R ¹ is selected from the group consisting of a to k;

a. NR ^a R ^b (wherein R ^a is and R ^b is independently hydrogen, C _{1 -4} alkyl, or (CH _{2) d} F ^18, d is an integer of 1 to 4),

b. Hydroxy,

c. C _{1 - 4} alkoxy,

d. Hydroxy (C _1-4 ) alkyl,

e. halogen,

f. Cyano,

g. Hydrogen,

h. Nitro,

i. (C ₁ -C ₄ ) alkyl,

j. Halo (C ₁ -C ₄ ) alkyl, and

k. Formyl;

R ^{1 '} is selected from the group consisting of a to f;

a. ¹²³ I, ¹²⁵ I, ¹³¹ I, ¹⁸ F or ⁷⁶ Br,

b. Hydrogen,

c. ¹⁸ F (C _{1 -4)} alkyl,

d. ^[18 F (C _{1 -4)} alkyl] amino,

e. [ ¹⁸ F (C ₁ -C ₄ ) alkyl] alkylamino, and

f. ¹⁸ F (C ₁ -C ₄ ) alkoxy;

R ² is selected from the group consisting of i to iv;

i. Hydroxyl, C _{1 - 4} alkoxy, (C ₁ -C ₄₎ - alkyl-oxo-alk (C ₁ -C ₄₎ aryloxy, (C ₁ -C ₄₎ - alkyl, oxo (C ₁ -C ₄₎ - alkyl, oxo ( C ₁ -C ₄ ) alkoxy, (C ₁ -C ₄ ) -alkyloxo (C ₁ -C ₄ ) -alkyloxo (C ₁ -C ₄ ) -alkyloxo (C ₁ -C ₄ ) alkoxy, carboxy (C ₁ -C ₄ ) alkyl, halo (C ₁ -C ₄ ) alkoxy, halo (C ₁ -C ₄ ) -alkyloxo (C ₁ -C ₄ ) alkoxy, halo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) -alkyloxy, halo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxy, halo (C ₁ -C ₄ ) alkyl, NR ⁶ R ^{6 ′} Wherein R ⁶ and R ^{6 ′} are independently selected from the group consisting of hydrogen, hydroxy (C ₁ -C ₄ ) alkyl and C ₁ -C ₄ alkyl), phenyl (C ₁ -C ₄ ) alkyl, ¹⁸ F ( C ₁ -C ₄ ) alkoxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkoxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C _1- C ₄ ) alkyloxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxy or ¹⁸ F (C ₁ -C ₄ ) alkyl;

ii.

Wherein q is an integer from 1 to 10; Z is ¹⁸ F, ¹⁸ F substituted benzoyloxy, ¹⁸ F substituted benzyloxy, preferably ¹⁸ F-phenoxy, ¹⁸ F substituted phenyl (C _{1- 4)} alkyl, ¹⁸ F-substituted (C _{1 -4)} alkoxy, ¹⁸ F and ¹⁸ F substituted aryloxy-substituted C _{6 -10} aryl, preferably is selected from the group consisting of ¹⁸ F- phenyl; R ^30, R ^31, R ³² and R ³³ are selected from hydrogen, hydroxy, C _{1 -4} alkoxy, C _{1 -4} alkyl and hydroxy (C _1-4) alkyl group consisting of independently, in each case.)

iii.

(Wherein Z, R ³⁰ , R ³¹ , R ³² and R ³³ are as described above).

iv.

Wherein Y is ¹⁸ F, ¹⁸ F substituted benzoyloxy, ¹⁸ F substituted phenyl (C _1-4 ) alkyl, ¹⁸ F substituted aryloxy, preferably ¹⁸ F-phenoxy and ¹⁸ F substituted C _6-10 aryl, and preferably is selected from the group consisting of ¹⁸ F- phenyl;

U is hydrogen, hydroxy, ¹⁸ F, ¹⁸ F substituted benzoyloxy, ¹⁸ F substituted phenyl (C _1-4 ) alkyl, ¹⁸ F substituted aryloxy, preferably ¹⁸ F-phenoxy and ¹⁸ F substituted C _{6 -10} aryl, preferably selected from the group consisting of ¹⁸ F- phenyl;

^{R 34, R 35, R 36} , R 37, R 38, R 39 and R ⁴⁰ are independently at each occurrence hydrogen, hydroxy, C _1-4 alkoxy, C _{1 -4} alkyl and hydroxy (C _1-4 ) Is selected from the group consisting of alkyl);

R ⁷ and R ⁸ are independently at each occurrence a halogen, e.g., F, Cl and Br, hydrogen, hydroxy, amino, methylamino, dimethylamino, C _{1 -4} alkoxy, C _{1 -4} alkyl and hydroxyl when (C _{1 - 4)} is selected from the group consisting of alkyl, wherein at least one of R ⁷ and R ⁸ is a halogen, preferably F.

In a preferred embodiment,

R ¹ is independently hydrogen, halogen, eg, F, Cl and Br, C ₁ -C ₄ alkyl, cyano, hydroxyl, nitro, (C ₁ -C ₄ ) alkylamino, di (C ₁ -C ₄ ) alkylamino, halo (C ₁ -C ₄ ) alkyl, formyl and alk (C ₁ -C ₄ ) -oxy;

R ^{1 'are} independently ^{hydrogen, 123 I, 125 I, 131} I, 18 F, 18 F (C 1 -4) _{^{alkyl, [18 F (C 1 -4}} ) alkyl] amino, ^[18 F (C _{1 - 4} ) alkyl] alkylamino, ¹⁸ F (C ₁ -C ₄ ) alkoxy and ⁷⁶ Br;

R ² is independently a hydroxyl, C _{1 - 4} alkoxy, (C ₁ -C ₄₎ alkyl alk-oxo (C ₁ -C ₄₎ alkyloxy, carboxy (C ₁ -C ₄₎ alkyl, halo (C ₁ -C ₄ ) -Alkoxy, halo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkoxy, halo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) -alkyloxo (C ₁ -C ₄ ) Alkyloxy, halo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo- (C ₁ -C ₄ ) alkyloxy, halo (C ₁ -C ₄ ) Alkyl, NR ⁶ R ^{6 ′} , wherein R ⁶ and R ^{6 ′} are independently selected from the group consisting of hydrogen, hydroxy (C ₁ -C ₄ ) alkyl and C ₁ -C ₄ alkyl, phenyl (C _1- C ₄ ) alkyl, ¹⁸ F (C ₁ -C ₄ ) alkoxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo- (C ₁ -C ₄ ) alkoxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo- ( C ₁ -C ₄ ) -alkyloxo (C ₁ -C ₄ ) alkyloxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo- ( C ₁ -C ₄ ) alkyloxy and ¹⁸ F (C ₁ -C ₄ ) alkyl;

R ⁷ and R ⁸ are selected from H, F, Cl or Br, wherein R ⁷ or R ⁸ is halogen.

In a more preferred embodiment,

R ¹ is selected from the group consisting of hydrogen, hydroxy, (C ₁ -C ₄ ) alkylamino, di (C ₁ -C ₄ ) alkylamino, methyl and methoxy, in particular hydrogen, methylamino and dimethylamino;

R ^{1 '} is selected from the group consisting of hydrogen, ¹²³ I, ¹²⁵ I, ¹³¹ I and ¹⁸ F, in particular hydrogen;

R ² is hydroxy, C _{1 - 4} alkoxy, NR ⁶ R ^{6 '} Wherein R ⁶ and R ^{6 ′} are independently selected from the group consisting of hydrogen, hydroxy (C ₁ -C ₄ ) alkyl and C ₁ -C ₄ alkyl), ¹⁸ F (C ₁ -C ₄ ) alkoxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkoxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxy and ¹⁸ F (C ₁ -C ₄ ) alkyl Group, in particular, (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkoxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyl Oxy and ¹⁸ F (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxy;

R ⁷ and R ⁸ are selected from the group consisting of hydrogen and fluorine, wherein R ⁷ or R ⁸ is fluorine.

Surprisingly, it has been found that stilbene derivatives with additional halogens, in particular fluorine atoms, to double bonds exhibit improved pharmacokinetic properties and / or increased metabolic stability and / or increased geometric isomer stability and uniformity.

Preferred compounds of formula (I) have the structure

Wherein one of R ⁷ and R ⁸ is hydrogen and the other is halogen.

A second aspect of the invention relates to a compound of formula II or a pharmaceutically acceptable salt thereof.

Where

R ¹ is selected from the group consisting of a to d;

a. NR ^a R ^b (where, R ^a and R ^b are independently hydrogen, C _{1 -4} alkyl or ^{_{(CH 2) d 18 F,}} d is an integer from 1 to 4, R ^a and R ^b both are oxygen And form nitro),

b. Hydroxy,

c. C _{1 - 4} alkoxy, and

d. Hydroxy (C _1-4 ) alkyl;

R ² is selected from the group consisting of i to iii;

i.

(Wherein, q is an integer of 1 to 10; Z is ¹⁸ F, ¹⁸ F-substituted benzoyloxy, ¹⁸ F-substituted (C _{1 -4)} alkoxy, ¹⁸ F-substituted benzyloxy, preferably ¹⁸ F- phenoxy, ¹⁸ F-substituted phenyl (C _1-4) alkyl, substituted aryloxy, and ¹⁸ F ¹⁸ F-substituted C _{6 -10} aryl, preferably is selected from the group consisting of ¹⁸ F- phenyl; R ^30, R ^31, R ³² and R ³³ are selected from hydrogen, hydroxy, C _{1 -4} alkoxy, C _{1 -4} alkyl and hydroxy (C _1-4) alkyl group consisting of independently, in each case.)

ii.

(Wherein Z, R ³⁰ , R ³¹ , R ³² and R ³³ are as described above).

iii.

Wherein Y is ¹⁸ F, ¹⁸ F substituted benzoyloxy, ¹⁸ F substituted phenyl (C _1-4 ) alkyl, ¹⁸ F substituted aryloxy, preferably ¹⁸ F-phenoxy and ¹⁸ F substituted C _6-10 aryl, and preferably is selected from the group consisting of ¹⁸ F- phenyl;

U is hydrogen, hydroxy, ¹⁸ F, ¹⁸ F substituted benzoyloxy, ¹⁸ F substituted phenyl (C _1-4 ) alkyl, ¹¹⁸ F substituted aryloxy, preferably ¹⁸ F-phenoxy and ¹⁸ F substituted C _{6 -10} aryl, preferably selected from the group consisting of ¹⁸ F- phenyl;

^{R 34, R 35, R 36} , R 37, R 38, R 39 and R ⁴⁰ are independently at each occurrence hydrogen, hydroxy, C _{1 -4} alkoxy, C _{1 -4} alkyl and hydroxy (C _1-4 ) Is selected from the group consisting of alkyl);

R ⁷ and R ⁸ are independently selected from hydrogen, hydroxy, amino, methylamino, dimethylamino, C _{1 -4} alkoxy, C _{1 -4} alkyl and hydroxy (C _1-4) alkyl at each occurrence the group consisting of do.

More preferably, the values of R ³⁰ , R ³¹ , R ³² , R ³³ , R ³⁴ , R ³⁵ , R ³⁶ , R ³⁷ , R ³⁸ , R ³⁹ and R ⁴⁰ are each independently hydrogen, hydroxy in each case , Amino, methylamino, dimethylamino and methoxy.

Preferably, R ² is meta or para position relative to the ethylene bridge.

When R ² is a group of the formula i, preferred values of R ³⁰ , R ³¹ , R ³² and R ³³ are hydrogen in each case and Z is ¹⁸ F. Useful values of q are integers from 1 to 10. Preferably, q is an integer from 2 to 5. More preferably, the value of q is 3 or 4.

i.

Preferred embodiments of compounds of formula (II) include the following structures wherein R ^a and R ^b are independently hydrogen or methyl, and preferably at least one of R ^a and R ^b is methyl.

Wherein X is ¹⁸ F.

A preferred series of compounds of formula (II) include ¹⁸ F labeled polyethylene glycol (PEG)-stilbene derivatives having the structure:

Wherein q is an integer from 1 to 10. More preferred compounds include those in which q is 2, 3 or 4, respectively, as follows.

In this series of compounds, ¹⁸ F is linked to stilbene through PEG chains having various numbers of ethoxy groups. All stilbene fluorides showed high binding affinity in assays with post AD brain homogenate (K _i = 2.9-6.7 nM). As can be seen in Schemes 1 to 3 herein, radiolabeling involves the substitution of 10a-d mesylate groups with [ ¹⁸ F] fluoride to target compound [ ¹⁸ F] 12a-d (EOS, specific activity, 900-1,500 Successful by providing Ci / mmol; radioactive compound purity> 99%). In vivo biodistribution of these ¹⁸ F ligands in normal mice shows good brain infiltration and rapid washout after intravenous injection (6.6-8.1 and 1.2-2.6% dose / g at 2 and 60 minutes, respectively). The specific binding related to the presence of Aβ plaques was confirmed by autoradiography of post-AD brain sections of [ ¹⁸ F] 12a-d . In vivo plaque labeling was also clearly demonstrated using these ¹⁸ F labeled agents in transgenic mice (Tg2576), a useful animal model for Alzheimer's disease.

The present invention also relates to compounds of the formula III.

Wherein n is an integer from 1 to 4, R ⁷ and R ⁸ are as described above, R ⁴¹ is selected from the group consisting of hydroxy and NR ^a R ^b , wherein R ^a and R ^b are independent is hydrogen or C _{1 -4} alkyl, R ^a and R ^b are both oxygen to form nitro.

Preferably, n is 1 and R ⁴¹ is hydroxy, methylamino or dimethylamino.

Under the preferred range of C _{6 -10} aryl comprises a phenyl, naphthyl or tetrahydro-naphthyl. Preferred under the scope of heteroaryl include thienyl, furyl, pyranyl, pyrrolyl, pyridinyl, indolyl and imidazolyl. Preferred within the scope of the heterocycle include piperidinyl, pyrrolidinyl and morpholinyl.

The compounds of the formulas (I), (II) and (III) can also be solvated, especially hydrated. Hydration may occur during the preparation of the compound or composition comprising the compound, or may occur over time due to the absorption properties of the compound. In addition, the compounds of the present invention may exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol and the like. In general, the solvated forms are considered equivalent to the unsolvated forms for the purposes of the present invention.

It is to be understood that the present invention includes stereoisomers of stilbene-based compounds, such as both cis and trans isomers. Further included are optical isomers, such as individual enantiomers and diastereomers as well as mixtures of enantiomers, which arise as a result of the structural asymmetry of the selected compound of formula (I), (II) or (III).

If any component or any variable in formula (I), (II) or (III) appears one or more times, its definition at each occurrence is independent of its definition at all other occurrences. In addition, combinations of substituents and / or variables may be acceptable only if such combinations result in stable compounds.

As used herein, alone or as part of another group, the term "alkyl" refers to both straight and branched chain radicals of 8 or less, preferably 6 or less, more preferably 4 or less, such as methyl, ethyl, propyl, Isopropyl, butyl, t-butyl and isobutyl.

The term "alkoxy" is used herein to mean a straight or branched chain alkyl radical, as defined above, attached to an oxygen atom (the chain length is not limited thereto) and is used to represent methoxy, ethoxy, n-pro Foxy, isopropoxy, and the like, but are not limited thereto. Preferably the alkoxy chain has 1 to 6 carbon atoms in length, more preferably 1 to 4 carbon atoms in length.

As used herein, alone or as part of another group, the term “monoalkylamine” means an amino group substituted with one alkyl group as defined above.

As used herein, alone or as part of another group, the term “dialkylamine” refers to an amino group substituted with two alkyl groups as defined above.

As used herein, alone or as part of another group, the terms "halo" or "halogen" refer to chlorine, bromine, fluorine or iodine unless specific use is otherwise defined in the text and / or claims.

The term "haloalkyl" as used herein refers to any alkyl group as described above substituted with one or more chlorine, bromine, fluorine or iodine, preferably fluorine or chlorine such as chloromethyl, iodomethyl, trifluoromethyl , 2,2,2-trifluoroethyl and 2-chloroethyl.

As used herein, alone or as part of another group, the term "aryl" refers to a monocyclic or bicyclic aromatic group containing 6 to 12 carbons in the ring position, preferably 6 to 10 carbons in the ring position, such as , Phenyl, naphthyl or tetrahydronaphthyl.

As used herein, the term "heterocycle" or "heterocyclic ring", except where indicated, may be saturated or unsaturated, and represent carbon atoms, and N, O and S (here nitrogen and sulfur heteroatoms). Represents a stable 5-7 membered mono-heterocyclic ring system composed of 1 to 3 heteroatoms selected from the group consisting of (optionally oxidizable). Particularly useful are rings containing one nitrogen atom or two nitrogen hetero atoms combined with one oxygen or sulfur. Examples of such heterocyclic groups are piperidinyl, pyrrolyl, pyrrolidinyl, imidazolyl, imidazinyl, imidazolidinyl, pyridyl, pyrazinyl, pyrimidinyl, oxazolyl, oxazolidinyl, isox Sazolyl, isoxazolidinyl, thiazolyl, thiazolidinyl, isothiazolyl, homopiperidinyl, homopiperazinyl, pyridazinyl, pyrazolyl and pyrazolidinyl, most preferably thiamorpholinyl, pi Ferrazinyl and morpholinyl.

The term "heteroatom" is used herein to mean an oxygen atom ("0"), a sulfur atom ("S") or a nitrogen atom ("N"). If the hetero atom is nitrogen, which R ^a and R ^b are each independently hydrogen, C _{1 -4} alkyl, C _{2 -4-amino-alkyl,} C _{1 -4} haloalkyl, or halo benzyl, NR ^a R ^b form a moiety, or or R ¹ and R ² are optionally recognize that O, S or NR ^c in the ring (wherein, R ^c is hydrogen or c _{1 -4} alkyl) having a 5 to 7 may form a heterocyclic ring will be.

As used herein, the term "heteroaryl" refers to 5 to 14 ring atoms; And a group having 6, 10 or 14 π electrons shared in a periodic arrangement and containing carbon atoms and 1, 2, 3 or 4 oxygen, nitrogen or sulfur heteroatoms, wherein examples of heteroaryl groups include Thienyl, benzo [b] thienyl, naphtho [2,3-b] thienyl, thianthrenyl, furyl, pyranyl, isobenzofuranyl, benzoxazolyl, chromenyl, xanthenyl, phenoxatii Nyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolinyl, isoindoleyl, 3H-indolyl, indolyl, indazolyl, Furinyl, 4H-quinolinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinazolinyl, cynolinyl, pterridinyl, 4aH-carbazolyl, carbazolyl, jab-carbolyl Nil, phenanthridyl, acridinyl, perimidinyl, phenanthrolinyl, phenazinyl, isothiazolyl, phenothiazinyl, isoxazolyl, furazanyl and phenoxazinyl groups have.

Herein, the term "aralkyl" or "arylalkyl" used alone or as part of another group is a C ₁ as described above having an aryl _{substituent-6} alkyl group, e.g., benzyl, phenylethyl or 2-naphthylmethyl Means.

The invention further relates to a process for the preparation of compounds of formula (I), (II) or (III). The compounds of the present invention can be prepared by the reactions described in Schemes 1-8 below.

Scheme 1 depicts a synthetic route for thiophene containing derivatives of formula (I), particularly for certain compounds of formula (Ia).

Fluorinated PEG stilbenes 12a-d are prepared by the reaction shown in Scheme 1. In order to prepare compounds having 2 or 3 ethoxy groups by PEG linkage, commercially available chlorides 2a, b can be coupled with OH groups of 4-methylamino-4'-hydroxy stilbene 1 (Ono M, et. al., Nucl Med Biol. 2003; 30: 565-71; Wilson A, et al., J Labelled Cpd Radiopharm. 2003; 46: S61), respectively, 3a, b . Free OH groups of 3a, b were then protected with TBDMSCl to afford compounds 7a, b . In order to prepare compounds having 4 or 5 ethoxy groups by PEG linkage, bromide 6c, d was separately prepared as shown in Scheme 2 and then coupled with stilbene 1 to obtain TBS protected compound 7c, d . Treatment with TBAF (1M) in THF removed the O-TBS protecting group on compounds 7c , d to afford 3c, d . The methylamino group of 7a-d was protected with BOC to give compound 8a-d . TBAF (1M) / THF was used to remove the 8a-d TBS protecting group and then reacted with MsCl in the presence of triethylamine to convert free OH groups to mesylate to afford 10a-d . 10a-d was refluxed in anhydrous TBAF / THF (Cox DP, et al., J Org Chem. 1984; 49: 3216-19) and then stirred with TFA to remove the BOC protecting group, thereby "cooling" fluorinated PEG stilbene 12a-d was obtained successfully.

To prepare the desired ¹⁸ F labeled PEG stilbene [ ¹⁸ F] 12a-d , N-BOC protected mesylate 10a-d was used as a precursor (Scheme 3). Each mesylate 10a-d was mixed with [ ¹⁸ F] fluoride / potassium carbonate and Kryptofix 222 in DMSO and heated at 120 ° C. for 4 minutes. The mixture was then treated with aqueous HCl solution to remove the N-BOC protecting group. The crude product was purified by HPLC (radioactive compound purity> 99%, radioactive compound yield 10-30%, decay corrected). The preparation of each ¹⁸ F labeled compound [ ¹⁸ F] 12a-d took about 90 minutes and the specific activity at the end of the synthesis was estimated to be 900-1,500 Ci / mmol.

Synthesis of compounds 15e, 16e and radiolabeled precursors 15d, 17d for preparing [ ¹⁸ F] 15e and [ ¹⁸ F] 16e are shown in Scheme 4. To prepare compound 15a , the nitro group of 4-nitro-4'-hydroxy stilbene 13a was reduced with SnCl ₂ in ethanol to give the corresponding amine 14a . The amino group was then treated with (CHO) _n and NaBH ₃ CN to give dimethylamino compound 15a . Hydroxy stilbene 15a was reacted with 20m bromide (prepared separately as shown in Scheme 5) and potassium carbonate in anhydrous DMF to give compound 15b . 15b was treated with 1N HCl in acetone to give compound 15c . Obtained. Mono tosylate 15d can be isolated from the product mixture of diol 15c and 1.5 equivalents of tosyl chloride in pyridine. Tosylate 15d was converted to fluoride 15e by refluxing with dry TBAF in THF. TBAF should be dried at 58 ° C. under high vacuum (<0.5 mmHg) for 24 hours prior to use. Tosyl compound 15d was used as starting material to obtain radiolabeled compound [ ¹⁸ F] 15e . Nitro compound 13e was similarly synthesized by tosylation and fluorination after coupling reaction of 13a and 20m . Synthesis of compound 16a was accomplished by reducing the nitro group of 13e with SnCl ₂ / EtOH and then monomethylating the amino group with (CHO) _n , NaOCH ₃ and NaBH ₄ . Intermediate 13b was reduced with amine 14c and then monomethylated to give compound 16c . To obtain [ ¹⁸ F] 16e , N-protected tosylate 17d was designed as precursor for radiolabeling. The previously prepared 14a was first monomethylated to 16a . Then 16a and 2 On Compound 17f was prepared by coupling of Scheme 5 and introducing BOC into the secondary amine. 17f of di-tert-butyl silyl group was removed at room temperature with 1N TBAF in THF to give diol 5c , which was monotosylated to give compound 17d .

Related compound 15h was also synthesized as shown in Scheme 6. Substituted malonate 21 was reduced to diol 22 using DIBALH and then reacted with 1 equivalent of TBSCl to give 23 . Unprotected OH was then converted to bromide 24 using CBr ₄ / PPh ₃ . Is reacted with 15a to compound 24 to yield the 15g, 15h was obtained by treating them with TBAF to remove TBS group.

In addition, two benzyl derivatives 14 and 15 of N, N-dimethyl stilbene were synthesized (Scheme 4). Compound 14 was obtained by reducing the corresponding ethyl ester 13 ³ with LiAlH ₄ . Benzyl alcohol was then converted to the highly reactive benzyl bromide intermediate with HBr / HOAc and converted directly to methyl ether 15 by addition of methanol and potassium carbonate without purification.

These stilbene derivatives in which the fluorine atom is directly attached to the double bond (formula I: R ⁷ or R ⁸ fluorine) were synthesized by well known methods (eg Tetrahedron Lett. 43, (2002), 2877- 2879).

To obtain [ ¹⁸ F] 15e , precursor 15d was mixed with [ ¹⁸ F] fluoride / potassium carbonate and Cryptopix® 222 in DMSO and heated at 120 ° C. for 4 minutes. The crude product was purified by HPLC to yield greater than 99% radioactive compound purity and 10% radioactive compound yield (compound decay corrected). The process took 90 minutes and the specific activity at the end of the synthesis was estimated at 70 Ci / mmol. Similar procedures were carried out to obtain [ ¹⁸ F] 16e from precursor 17d . After the initial reaction in DMSO, the mixture was treated with aqueous HCl solution to remove the BOC groups. After HPLC purification the radioactive compound purity was greater than 99% and the radioactive compound yield was 15%. The total synthesis took 110 minutes and the specific reactivity at the end of the synthesis was estimated to be 90 Ci / mmol.

Some of the compounds are modifiable by microwave synthesis as described in Examples 50-52 below.

The radiohalogenated compounds of the invention can be readily formed from materials that can be provided in a kit. Kits for forming imaging agents may include, for example, vials containing a physiologically suitable solution of the intermediate of formula (I) at a concentration and pH suitable for optimal complexing conditions. The user will add an appropriate amount of radioisotope and oxidant such as hydrogen peroxide to the vial. The resulting labeled ligand is then administered intravenously to the patient and receptors in the brain will be imaged by measuring gamma rays or light emission therefrom.

If desired, the radiodiagnostic agent may contain any additives such as pH adjusting agents (eg, acids, bases, buffers), stabilizers (eg, ascorbic acid) or isotonic agents (eg, sodium chloride). Can be.

As used herein, the term “pharmaceutically acceptable salts” is within the scope of sound medical judgment, free of inadequate toxicity, irritation, allergic reactions, etc., having a reasonable benefit / risk ratio, effective for the intended use, and effective for patients. By carboxylic acid salt or acid addition salt of a compound of the invention, suitable for use in contact with tissue, it means a zwitterionic form of a compound of the invention, if possible. The term "salt" means a relatively nontoxic inorganic and organic acid addition salt of a compound of the invention. Also included are aliphatic mono and dicarboxylic acids, for example derived from acetic acid, phenyl-substituted alkanoic acid, hydroxy alkanoic acid and alkanedic acid, aromatic acids, and nontoxic organic acids such as aliphatic and aromatic sulfonic acids. These salts. These salts can be prepared in situ during the final isolation and purification of the compound or by reacting the free base form of the purified compound separately with the appropriate organic or inorganic acid and isolating the salt so formed. Further representative salts are hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate , Tosylate, citrate, maleate, fumarate, succinate, tartarate, naphthylate, mesylate, glucoheptonate, lacthiobionate and laurylsulfonate salts, propionate, pivalate, sicle Lamate, isethionate, and the like. These are cations based on alkali and alkaline earth metals such as sodium, lithium, potassium, calcium, magnesium and the like, and ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine Non-toxic ammonium, quaternary ammonium and amine cations, including but not limited to, and the like (see, eg, Berge SM, et al., Pharmaceutical Salts, J. Pharm. Sci, incorporated herein by reference). 66: 1-19 (1977).

The invention also relates to a method of imaging amyloid deposits. One of the important preliminary requirements for in vivo imaging of the brain is the ability to cross the intact blood-brain barrier after intravenous bolus injection.

In the first step of the imaging method of the invention, a detectable amount of a labeled compound of Formula (I), (II) or (III) is introduced into a tissue or patient. The compound is typically part of a pharmaceutical composition and is administered to a tissue or patient by methods well known to those skilled in the art.

For example, the compound may be administered orally, rectally, parenterally (intravenously, intramuscularly or subcutaneously), in tank, vaginal, intraperitoneal, in bladder, topical (powder, ointment or drip) or in intraoral or intranasal sprays. Can be.

In a preferred embodiment of the invention, the labeled compound is introduced into the patient in a detectable amount, and after sufficient time for the compound to bind the amyloid deposit, the labeled compound is detected non-invasively in the patient. In another embodiment of the invention, an ¹⁸ F labeled compound of Formula (I), (II) or (III) is introduced into a patient and allowed sufficient time for the compound to bind to an amyloid deposit, after which the tissue sample of the patient is isolated to Labeled compounds in are detected away from the patient. In a third embodiment of the invention, a tissue sample is separated from a patient to introduce a labeled compound of formula (I), (II) or (III) into the tissue sample. After sufficient time has passed for the compound to bind with the amyloid deposit, the compound is detected.

The labeled compound can be administered to the patient in a general or topical route of administration. For example, the labeled compound can be administered to a patient for delivery systemically. Alternatively, the labeled compound can be administered to certain subject organs or tissues. For example, it is desirable to locate and quantify amyloid deposits in the brain to diagnose or track the progression of Alzheimer's disease in a patient.

The term "tissue" refers to a part of the patient's body. Examples of tissues include brain, heart, liver, blood vessels and arteries. The detectable amount is the amount of labeled compound required to be detected by the selected detection method. The amount of labeled compound introduced into the patient to be detected can be readily determined by one skilled in the art. For example, an increased amount of labeled compound can be administered to a patient until the compound is detected by the selected detection method. The label is introduced into the compound to provide detection of the compound.

The term "patient" means a human or other animal. Those skilled in the art are also familiar with determining the amount of time sufficient for a compound to bind an amyloid deposit. The amount of time required can be readily determined by introducing a detectable amount of a labeled compound of Formula (I), (II) or (III) into the patient and then detecting the labeled compound at various time points after administration.

The term "associated" means chemical interaction between the labeled compound and amyloid deposits. Examples of bonds include covalent bonds, ionic bonds, hydrophilic-hydrophilic interactions, hydrophobic-hydrophobic interactions and complexes.

Those skilled in the art are familiar with proton-emitting tomography (PET) detection of proton-emitting atoms such as ¹⁸ F. The invention also relates to certain compounds in which ¹⁸ F atoms have been replaced with non-radiolabeled fluorine atoms.

The radiodiagnostic agent should have sufficient radiation and radioactivity concentrations to ensure a reliable diagnosis. Desired levels of radioactivity can be achieved by the methods provided herein for the preparation of compounds of Formula (I), (II) or (III).

Imaging of amyloid deposits can be performed quantitatively such that the amount of amyloid deposits can be determined.

Another aspect of the invention is a method of inhibiting amyloid plaque aggregation. The present invention also provides a method of inhibiting amyloid protein aggregation and formation of amyloid deposits by administering to the patient an amyloid inhibitory amount of the compound of formula (I), (II) or (III).

One skilled in the art can easily determine the amount of amyloid inhibition by simply administering to the patient an increasing amount of a compound of Formula I, II or III until the growth of amyloid deposits is reduced or stopped. Growth rates can be assessed using imaging as described above, or by taking a tissue sample from a patient and observing amyloid deposits therein. Compounds of the invention may be administered to a patient at a dosage level in the range of about 0.1 to about 1,000 mg per day. For a normal human adult having a weight of about 70 kg, a dose in the range of about 0.01 to about 100 mg per kg body weight per day is sufficient. However, the specific dose used may vary. For example, the dosage may vary depending on a number of factors, including the requirements of the patient, the severity of the disease to be treated and the pharmacological activity of the compound used. Determination of the optimal dose for a particular patient is well known to those skilled in the art.

The radiodiagnostic agent should have sufficient radiation and radioactivity concentrations to ensure a reliable diagnosis. Desired levels of radioactivity can be achieved by the methods provided herein for the preparation of compounds of Formula (I), (II) or (III).

In addition, imaging of amyloid deposits can be performed quantitatively such that the amount of amyloid deposits can be determined.

The following examples illustrate, but do not limit, the methods and compositions of the present invention. Other suitable modifications and adjustments of the various conditions and parameters commonly encountered by those skilled in the art and apparent to those skilled in the art are within the spirit and scope of the present invention.

All reagents used for the synthesis are commercial products and used without further purification unless otherwise specified. ¹ H NMR spectra were obtained on a Bruker DPX spectrometer (200 MHz) in CDCl ₃ . Chemical shifts are reported as δ values (parts per million) for internal TMS. Coupling constants are reported in hertz. Multiplicity is defined as s (single), d (double), t (triple), br (wide), m (multiple). Elemental analysis was performed by Atlantic Microlab INC. For each process, "standard workup" means the following steps: addition of the specified organic solvent, washing the organic layer with water, then brine, separating the organic layer from the aqueous layer, drying the combined organic layers with anhydrous sodium sulfate and filtering sodium sulfate Removal and removal of the organic solvent under reduced pressure.

Example  One

2- (2- {4- [2- (4-Methylamino-phenyl) -vinyl] -phenoxy} -ethoxy) -ethanol ( 3a ).

Under a nitrogen atmosphere, 4-methylamino-4'-hydroxy stilbene 1 (Ono M, et al., Nucl Med Biol. 2003; Wilson A, et al., J Labelled Cpd Radiopharm. 2003) (63 mg, 0.28 mmol) and 2a (42 mg, 0.34 mmol) were dissolved in anhydrous DMF (5.0 ml) and potassium carbonate (125 mg, 0.91 mmol) was added. The suspension was heated to 100 ° C. and stirred overnight. After cooling to room temperature, standard workup was applied with dichloromethane and the residue was purified by TLC for silica gel preparation (4% methanol in dichloromethane) to give compound 3a (67 mg, 76%).

Example  2

2-[-2- (2- {4- [2- (4-Methylamino-phenyl) -vinyl] -phenoxy} -ethoxy) -ethoxy] -ethanol ( 3b ).

Compound 3b is prepared from 1 (150 mg, 0.67 mmol), 2b (136 mg, 0.81 mmol) and potassium carbonate (277 mg, 2.01 mmol) in DMF (10 ml) using the same method as described for compound 3a It was. 3b (180 mg, 76%):

Example 3

2- {2- [2- (2- {4- [2- (4-Methylamino-phenyl) -vinyl] -phenoxy} -ethoxy) -ethoxy] -ethoxy} -ethanol ( 3c ).

TBAF (1M in THF, 0.06 ml) was added via syringe to a solution of compound 7c (12 mg, 0.023 mmol) in THF (1 ml). The solution was stirred at rt for 2 h. After standard workup with dichloromethane, the residue was purified by TLC for silica gel preparation (4.5% methanol in dichloromethane) to give 3c (8.7 mg, 94%).

Example  4

2- (2- {2- [2- (2- {4- [2- (4-methylamino-phenyl) -vinyl] -phenoxy} -ethoxy) -ethoxy] -ethoxy} -ethoxy ) -Ethanol ( 3d ).

Compound 3d was prepared from 7AF (15 mg, 0.027 mmol) and TBAF in THF (1 ml, 1M in THF, 0.06 ml) using the same method as described for compound 3c . 3d (7.8 mg, 65%):

Example  5

2- (2- {2- [2- (tert-Butyl-dimethyl-silanyloxy) -ethoxy] -ethoxy} -ethoxy) -ethanol ( 5c ).

Tetraethylene glycol 4c (1.12 g, 5.77 mmol) and TBDMSCl (0.87 g, 5.77 mmol) were dissolved in dichloromethane (25 ml) followed by addition of triethyl amine (1.46 g, 14.4 mmol). The solution was stirred at rt for 2 h. After standard workup with dichloromethane, the residue was purified by silica gel column chromatography (50% ethyl acetate in hexanes) to give 5c (744 mg, 42%).

Example  6

2- [2- (2- {2- [2- (tert-butyl-dimethyl-silanyloxy) -ethoxy] -ethoxy} -ethoxy) -ethoxy] -ethanol ( 5d ).

Compound 5d was pentaethylene glycol 4d (1.13 g, 4.72 mmol), TBDMSCl (0.78 g, 5.19 mmol) and triethyl amine (1.2 g, in dichloromethane (25 ml) using the same method as described for compound 5c . 11.8 mmol). 5d (668 mg, 40%):

Example  7

(2- {2- [2- (2-Bromo-ethoxy) -ethoxy] -ethoxy} -ethoxy) -tert-butyl-dimethyl-silane ( 6c ).

Compound 5c (680 mg, 2.20 mmol) and carbon tetrabromide (947 mg, 2.86 mg) were dissolved in dichloromethane (20 ml). The solution was cooled to 0 ° C. using an ice bath, pyridine (2.0 ml) was added followed by triphenylphosphine (749 mg, 0.286 mmol). The solution was stirred at 0 ° C. for 30 minutes and then at room temperature for 2 hours. After standard workup with dichloromethane, the residue was purified by silica gel column chromatography (20% ethyl acetate in hexane) to give compound 6c (680 mg, 79.6%).

Example  8

[2- (2- {2- [2- (2-Bromoethoxy) -ethoxy] -ethoxy} -ethoxy) -ethoxy] -tert-butyl- dimethylsilane ( 6d ).

Compound 6d was purified using the same method as described for compound 6c , 5d (624 mg, 1.77 mmol), carbon tetrabromide (761 mg, 2.30 mmol), triphenylphosphine (602 mg, 2.30 mmol) and pyridine (2.0 ml). 6d (400 mg, 52.3%):

Example 9

{4- [2- (4- {2- [2- (tert-butyl-dimethyl-silanyloxy) -ethoxy] -ethoxy} -phenyl) -vinyl] -phenyl} -methyl-amine ( 7a ) .

Compound 3a (45 mg, 0.14 mmol) and TBDMSCl (33 mg, 0.22 mmol) were dissolved in dichloromethane (10 ml) and then imidazole (20 mg, 0.29 mmol) was added. The solution was stirred at rt for 2 h. After standard workup with dichloromethane, the residue was purified by silica gel column chromatography (1.5% methanol in dichloromethane) to give 7a (56 mg, 91%).

Example  10

(4- {2- [4- (2- {2- [2- (tert-butyl-dimethyl-silanyloxy) -ethoxy] -ethoxy} -ethoxy) -phenyl] -vinyl} -phenyl) -Methyl-amine ( 7b ).

Compound 7b was prepared from 3b (136 mg, 0.38 mmol), TBDMSCl (86 mg, 0.57 mmol) and imidazole (52 mg, 0.76 mmol) in dichloromethane (10 ml) using the same method as described for compound 7a . Prepared. 7b (170 mg, 95%):

Example 11

[4- (2- {4- [2- (2- {2- [2- (tert-butyl-dimethyl-silanyloxy) -ethoxy] -ethoxy} -ethoxy) -ethoxy] -phenyl } -Vinyl) -phenyl] -methyl-amine ( 7c ).

Compound 7c was subjected to the same method as described for compound 3a using 1 (98 mg, 0.44 mmol), 6c (210 mg, 0.57 mmol) and K ₂ CO ₃ (300 mg, 2.18 mmol) in DMF (10 ml). Prepared from. 7c (213 mg, 95%):

Example  12

{4- [2- (4- {2- [2- (2- {2- [2- (tert-butyl-dimethyl-silanyloxy) -ethoxy] -ethoxy} -ethoxy) -ethoxy ] -Ethoxy} -phenyl) -vinyl] -phenyl} -methyl-amine ( 7d ).

Compound 7d was used in the same manner as described for compound 3a using 1 (97 mg, 0.43 mmol), 6d (197 mg, 0.47 mmol) and K ₂ CO ₃ (297 mg, 2.15 mmol) in DMF (10 ml). Prepared from. 7d (220 mg, 91%):

Example  13

{4- [2- (4- {2- [2- (tert-butyl-dimethyl-silanyloxy) -ethoxy] -ethoxy} -phenyl) -vinyl] -phenyl} -methyl-carbamic acid tert- Butyl ester ( 8a ).

Under a nitrogen atmosphere, 7a (54 mg, 0.13 mmol) was dissolved in anhydrous DMF (5.0 ml) and then Boc-anhydride (84 mg, 0.25 mmol) was added. The solution was refluxed overnight. After standard workup with dichloromethane, the residue was purified by TLC for silica gel preparation (2% methanol in dichloromethane) to give 8a (60 mg, 90%).

Example  14

(4- {2- [4- (2- {2- [2- (tert-Butyl-dimethyl-silanyloxy) -ethoxy] -ethoxy} -ethoxy) -phenyl] -vinyl} -phenyl) -Methyl-carbamic acid tert-butyl ester ( 8b ).

Compound 8b was prepared from 7b (124 mg, 0.26 mmol) and Boc-anhydride (218 mg, 0.66 mmol) in THF (10 ml) using the same method as described for compound 8a . 8b (130 mg, 86%):

Example  15

[4- (2- {4- [2- (2- {2- [2- (tert-butyl-dimethyl-silanyloxy) -ethoxy] -ethoxy} -ethoxy) -ethoxy] -phenyl } -Vinyl) -phenyl] -methyl-carbamic acid tert-butyl ester ( 8c ).

Compound 8c was prepared from 7c (84 mg, 0.16 mmol) and Boc-anhydride (163 mg, 0.49 mmol) in THF (5 ml) using the same method as described for compound 8a . 8c (86 mg, 86%):

Example 16

{4- [2- (4- {2- [2- (2- {2- [2- (tert-butyl-dimethyl-silanyloxy) -ethoxy] -ethoxy} -ethoxy) -ethoxy ] -Ethoxy} -phenyl) -vinyl] -phenyl} -methyl-carbamic acid tert-butyl ester ( 8d ).

Compound 8d was prepared from 7d (210 mg, 0.51 mmol) and Boc-anhydride (840 mg, 2.54 mmol) in THF (10 ml) using the same method as described for compound 8a . 8d (174 mg, 66.7%):

Example 17

[4- (2- {4- [2- (2-hydroxy-ethoxy) -ethoxy] -phenyl} -vinyl) -phenyl] -methyl-carbamic acid tert-butyl ester ( 9a ).

Compound 9a was prepared from 8a (56 mg, 0.11 mmol) and TBAF (1M in THF, 0.21 ml) in THF (5 ml) using the same method as described for compound 3c . 9a (36 mg, 82%):

Example 18

{4- [2- (4- {2- [2- (2-hydroxy-ethoxy) -ethoxy] -ethoxy} -phenyl) -vinyl] -phenyl} -methyl-carbamic acid tert-butyl ester ( 9b ).

Compound 9b was prepared from 8b (118 mg, 0.21 mmol) and TBAF (1M in THF, 0.42 ml) in THF (10 ml) using the same method as described for compound 3c . 9b (94 mg, 99.7%):

Example 19

(4- {2- [4- (2- {2- [2- (2-hydroxy-ethoxy) -ethoxy] -ethoxy} -ethoxy) -phenyl] -vinyl} -phenyl) -methyl Carbamic acid tert-butyl ester ( 9c ).

Compound 9c was prepared from 8b (66 mg, 0.11 mmol), TBAF (1M in THF, 0.22 ml) and THF (5 ml) using the same method as described for compound 3c . 9c (50 mg, 93.0%):

Example 20

[4- (2- {4- [2- (2- {2- [2- (2-hydroxy-ethoxy) -ethoxy] -ethoxy} -ethoxy) -ethoxy] -phenyl}- Vinyl) -phenyl] -methyl-carbamic acid tert-butyl ester ( 9d ).

Compound 9d was prepared from 8d (76 mg, 0.12 mmol) and TBAF (1M in THF, 0.24 ml) in THF (5 ml) using the same method as described for compound 3c . 9d (52 mg, 82.7%):

Example  21

Methanesulfonic acid 2- [2- (4- {2- [4- (tert-butoxycarbonyl-methyl-amino) -phenyl] -vinyl} -phenoxy) -ethoxy] -ethyl ester ( 10a ).

Compound 9a (36 mg, 0.087 mmol) was dissolved in dichloromethane (5 ml) and triethylamine (44 mg, 0.44 mmol) was added. Methanesulfonyl chloride (30 mg, 0.26 mmol) was then added via syringe. The solution was stirred at rt for 4 h. After standard workup with dichloromethane, the residue was purified by TLC for silica gel preparation (2.0% methanol in dichloromethane) to give 10a (39 mg, 91%).

Example  22

Methanesulfonic acid 2- {2- [2- (4- {2- [4- (tert-butoxycarbonyl-methyl-amino) -phenyl] -vinyl} -phenoxy) -ethoxy] -ethoxy}- Ethyl ester ( 10b ).

Compound 10b was prepared using the same method as described for compound 10a , 9b (81 mg, 0.18 mmol), methanesulfonyl chloride (62 mg, 0.54 mmol) and triethylamine (88 mg, in dichloromethane (8 ml). 0.88 mmol). 10b (82 mg, 86.5%):

Example 23

Methanesulfonic acid 2- (2- {2- [2- (4- {2- [4- (tert-butoxycarbonyl-methyl-amino) -phenyl] -vinyl} -phenoxy) -ethoxy]- Methoxy} -ethoxy) -ethyl ester ( 10c ).

Compound 10c was purified using the same method as described for compound 10a , 9c (50 mg, 0.10 mmol), methanesulfonyl chloride (46 mg, 0.40 mmol) and triethylamine (50 mg, in dichloromethane (5 ml). 0.50 mmol). 10c (56 mg, 96.9%):

Example 24

Methanesulfonic acid 2- [2- (2- {2- [2- (4- {2- [4- (tert-butoxycarbonyl-methyl-amino) -phenyl] -vinyl} -phenoxy) -ethoxy ] -Ethoxy} -ethoxy] -ethoxy) -ethyl ester ( 10d ).

Compound 1 0d was converted to 9d (58 mg, 0.11 mmol), methanesulfonyl chloride (49 mg, 0.43 mmol) and triethylamine (54 mg) in dichloromethane (5 ml) using the same method as described for compound 10a . , 0.54 mmol). 10d (63 mg, 95%):

Example  25

[4- (2- {4- [2- (2-Fluoro-ethoxy) -ethoxy] -phenyl} -vinyl) -phenyl] -methyl-carbamic acid tert-butyl ester ( 11a ).

Anhydrous TBAF (Cox DP, et al., J Org Chem. 1984; 49: 3216-19) (38.5 mg, 0.15 mmol) was added to a solution of compound 10a (14.5 mg, 0.03 mmol) in anhydrous THF (3 ml). It was. The mixture was refluxed for 4 hours. After cooling to room temperature, standard workup with dichloromethane was carried out and the residue was purified by TLC for silica gel preparation (2% methanol in dichloromethane) to give compound 11a (7 mg, 57%).

Example 26

{4- [2- (4- {2- [2- (2-Fluoro-ethoxy) -ethoxy] -ethoxy} -phenyl) -vinyl] -phenyl} -methyl-carbamic acid tert-butyl ester ( 11b ).

Compound 11b was prepared from 10b (21 mg, 0.04 mmol) and TBAF (52 mg, 0.2 mmol) in THF (10 ml) using the same method as described for compound 11a . 11b (17 mg, 94%):

Example  27

(4- {2- [4- (2- {2- [2- (2-Fluoro-ethoxy) -ethoxy] -ethoxy} -ethoxy) -phenyl] -vinyl} -phenyl) -methyl -Carbamic acid tert-butyl ester ( 11c ).

Compound 11c was prepared from 10c (18 mg, 0.03 mmol) and TBAF (42 mg, 0.16 mmol) in THF (5 ml) using the same method as described for compound 11a . 11c (12 mg, 77%):

Example  28

[4- (2- {4- [2- (2- {2- [2- (2-Fluoro-ethoxy) -ethoxy] -ethoxy} -ethoxy) -ethoxy] -phenyl}- Vinyl) -phenyl] -methyl-carbamic acid tert-butyl ester ( 11d ).

Compound 11d was prepared from 10d (15 mg, 0.024 mmol) and TBAF (32 mg, 0.12 mmol) in THF (5.0 ml) using the same method as described for compound 11a . 11d (11 mg, 84%):

Example 29

[4- (2- {4- [2- (2-Fluoro-ethoxy) -ethoxy] -phenyl} -vinyl) -phenyl] -methyl-amine ( 12a ).

Trifluoroacetic acid (0.5 ml) was slowly added to a solution of compound 11a (7.0 mg, 0.017 mmol) in dichloromethane (1 ml). The mixture was then stirred at rt for 1 h. After standard workup with dichloromethane, the residue was purified by TLC for silica gel preparation (1.0% methanol in dichloromethane) to give compound 12a (3 mg, 56%).

Example  30

{4- [2- (4- {2- [2- (2-Fluoro-ethoxy) -ethoxy] -ethoxy} -phenyl) -vinyl] -phenyl} -methyl-amine ( 12b ).

Compound 12b was prepared from 11b (17 mg, 0.037 mmol) in trifluoroacetic acid (1 ml) and dichloromethane (2 ml) using the same method as described for compound 12a . 12b (9 mg, 68%):

Example 31

(4- {2- [4- (2- {2- [2- (2-Fluoro-ethoxy) -ethoxy] -ethoxy} -ethoxy) -phenyl] -vinyl} -phenyl) -methyl -Amine ( 12c ).

Compound 12c was prepared from 11c (12 mg, 0.024 mmol) in trifluoroacetic acid (0.5 ml) and dichloromethane (1 ml) using the same method as described for compound 12a . 12c (7 mg, 73%):

Example 32

[4- (2- {4- [2- (2- {2- [2- (2-Fluoro-ethoxy) -ethoxy] -ethoxy} -ethoxy) -ethoxy] -phenyl}- Vinyl) -phenyl] -methyl-amine ( 12d ).

Compound 12d was prepared from 11d (10 mg, 0.018 mmol) in trifluoroacetic acid (0.3 ml) and dichloromethane (1 ml) using the same method as described for compound 12a . 12d (6 mg, 73%):

Example 33

[ ¹⁸ F] [4- (2- {4- [2- (2-Fluoro-ethoxy) -ethoxy] -phenyl} -vinyl) -phenyl] -methyl-amine ([ ¹⁸ F] 12a ).

[ ¹⁸ F] fluoride prepared by cyclotron using an ¹⁸ O (p, n) ¹⁸ F reaction was prepared using a Sep-Pak Light QMA cartridge as an aqueous solution in [ ¹⁸ O] -rich water. Passed through. The cartridge was dried by airflow and the ¹⁸ F activity was monitored by Cryptofix 222 (K222) / K ₂ CO ₃ solution (22 mg K222 and 4.6 mg K ₂ CO ₃ ) 2 in CH ₃ CN / H ₂ O 1.77 / 0.23. eluted with mL. The solvent was removed at 120 ° C. under argon vapor. The residue was dried twice with 1 mL of anhydrous CH ₃ CN at 120 ° C. under argon vapor as a boiling mixture. A solution of mesylate precursor 10a (4 mg) in DMSO (0.2 mL) was added to the reaction vessel containing the dried ¹⁸ F active. The solution was heated at 120 ° C. for 4 minutes. Water (2 mL) was added and the solution was cooled for 1 minute. Then HCl (10% aqueous solution, 0.5 mL) was added and the mixture was again heated to 120 ° C. for 5 minutes. The pH was adjusted to basic (pH 8-9) by addition of aqueous NaOH solution. The mixture was extracted with ethyl acetate (1 mL × 2), the combined organic layers were dried (Na ₂ SO ₄ ) and the solvent removed using moderate heating (55-60 ° C.) under argon vapor. The residue was dissolved in CH ₃ CN and injected into HPLC for purification [Hamilton PRP-1 semi-prep column (7.0 × 305 mm, 10 μm), CH ₃ CN / dimethylglutarate buffer (5 mM , pH 7) 9/1; Flow rate 2 mL / min]. The retention time of 12a in this HPLC system was 8.9 minutes and was well separated from precursor 10a (hold time = 12 minutes) and hydrolysis byproducts (hold time = 6.2 minutes). The preparation took 90 minutes and the radioactive compound yield was 20% (disintegration corrected). To determine radioactive compound purity and specific activity, analytical HPLC was used [Hamilton PRP-1 analytical column (4.1 × 250 mm, 10 μm), CH ₃ CN / dimethylglutarate buffer (5 mM, pH 7 ) 9/1; Flow rate 0.5 mL / min]. The retention time of 12a in this system was 10.8 minutes and the radioactive purity (RCP) was over 99%. Specific activity was assessed by comparing the UV peak intensity of purified [ ¹⁸ F] 10 with known concentrations of the reference non-radioactive compound. The specific activity after preparation was 1,000-1,500 Ci / mmol.

Example  34

[ ¹⁸ F] {4- [2- (4- {2- [2- (2-Fluoro-ethoxy) -ethoxy] -ethoxy} -phenyl) -vinyl] -phenyl} -methyl-amine ( [ ¹⁸ F] 12b ).

[ ¹⁸ F] 12b was obtained from 10b using a similar reaction. The radioactive compound yield was 30% (decay corrected) and the radioactive compound purity exceeded 99%. The HPLC retention time of 12b for the analytical system described above was 11.7 minutes (specific activity = 1,300-1,500 Ci / mmol).

Example  35

[ ¹⁸ F] [(4- {2- [4- (2- {2- [2- (2-Fluoro-ethoxy) -ethoxy] -ethoxy} -ethoxy) -phenyl] -vinyl} -Phenyl) -methyl-amine ([ ¹⁸ F] 12c ).

[ ¹⁸ F] 12c was obtained from 10c using a similar reaction. The radioactive compound yield was 10% (decay corrected) and the radioactive compound purity exceeded 99%. HPLC retention time of 12c for the analytical system described above was 11.7 min (specific activity = 900 Ci / mmol).

Example  36

[ ¹⁸ F] [[4- (2- {4- [2- (2- {2- [2- (2-fluoro-ethoxy) -ethoxy] -ethoxy} -ethoxy) -ethoxy ] -Phenyl} -vinyl) -phenyl] -methyl-amine ([ ¹⁸ F] 12d ).

[ ¹⁸ F] 12d was obtained from 10b using a similar reaction. The radioactive compound yield was 20% (disintegration corrected) and the radioactive compound purity exceeded 99%. The HPLC retention time of 12d for the analytical system described above was 10.7 minutes (specific activity = 1,000-1,500 Ci / mmol).

Example  37

4-amino-4'-hydroxyl stilbene ( 14a )

First tin chloride (11.8 g, 0.062 mol) was added to a solution of compound 13a (Frinton Lab) (3.0 g, 0.012 mol) in ethanol (100 mL), followed by concentrated hydrochloric acid (5.0 mL). It was. The solution was refluxed for 3 hours and cooled to room temperature with stirring overnight. Aqueous sodium hydroxide solution (1N) was added to adjust the pH to 8.5-9. After standard workup with dichloromethane, crude product 14a was obtained (2.6 g, ˜100%). The product was used in the next step without further purification.

Example 38

4-N, N'-dimethylamino-4'-hydroxyl stilbene ( 15a )

To a mixture of 14a (211 mg, 1.0 mmol), paraformaldehyde (300 mg, 10 mmol) and sodium cyanoborohydride (189 mg, 3.0 mmol) was added acetic acid (10 mL). The whole mixture was stirred overnight at room temperature and then poured into 100 mL of water. Sodium carbonate was added to adjust the pH to 8-9. After standard workup with 5% methanol in dichloromethane, the residue was purified by silica gel column chromatography (2.5% methanol in dichloromethane) to give 15a as a white solid (214 mg, 89.5%).

Example  39

4-N, N'-dimethylamino-4 '-(2,2-dimethyl- [1,3] dioxan-5-ylmethoxy) stilbene ( 15b )

Under a nitrogen atmosphere, 15a (100 mg, 0.38 mmol) was dissolved in anhydrous DMF (5.0 mL). To this solution was added potassium carbonate (140 mg, 1.0 mmol), followed by 20m ¹ (105 mg, 0.5 mmol) of 5-bromomethyl-2,2-dimethyl- [1,3] dioxane. The mixture was heated to 100 ° C. and stirred overnight. After cooling to room temperature, standard workup with dichloromethane was carried out and the residue was purified by TLC for silica gel preparation (1% methanol in dichloromethane) to give compound 15b (100 mg, 72%).

Example  40

4-N, N'-dimethylamino-4 '-(1,3-dihydroxy-propan-2-ylmethoxy) stilbene ( 15c )

Compound 15b (180 mg, 0.49 mmol) was suspended in acetone (5.0 mL) and cooled to 0 ° C. using an ice bath. 1N HCl (5.0 mL, 5.0 mmol) was added slowly over 20 minutes. The suspension turned into a clear solution during the addition. The solution was stirred for an additional 30 minutes at 0 ° C. and then warmed to room temperature for 30 minutes. Saturated sodium bicarbonate solution was added to adjust the pH to 8.5-9. After standard workup with dichloromethane, the residue was purified by TLC for silica gel preparation (5% methanol in dichloromethane) to afford compound 15c as a white solid (140 mg, 87%).

Example  41

4-N, N'-dimethylamino-4 '-(1-tosyl-3-hydroxy-propan-2-ylmethoxy) stilbene ( 15d )

Compound 15c (158 mg, 0.49 mmol) was dissolved in anhydrous pyridine (15 mL) and cooled to 0 ° C. using an ice bath. Tosyl chloride (137 mg, 0.72 mmol) was added and the solution was stirred at 0 ° C. for 2 hours. After standard workup with dichloromethane, the residue was purified by TLC for silica gel preparation (5% methanol in dichloromethane) to give monotosylate compound 15d as a white solid (95 mg, 41%).

Example  42

4-N, N'-dimethylamino-4 '-(1-fluoro-3-hydroxy-propan-2-ylmethoxy) stilbene ( 15e )

Compound 15d (40 mg, 0.083 mmol) was dissolved in anhydrous THF (5 mL). Under nitrogen atmosphere, anhydrous TBAF (150 mg, 0.5 mmol) in anhydrous THF (1.0 mL) was added slowly. The solution was then heated to reflux for 3 hours. After cooling to room temperature, a standard workup with dichloromethane was carried out and the residue was subjected to silica gel preparation TLC (5% methanol in dichloromethane) to give product 15e (17 mg, 62%).

Example 43

4-nitro-4 '-(2,2-dimethyl- [1,3] dioxan-5-ylmethoxy) stilbene ( 13b )

Compound 13b was prepared from 13a (241 mg, 1.0 mmol) using the same method as described for compound 15b . 13b (260 mg, 70%):

Example  44

4-nitro-4 '-(1,3-dihydroxy-propan-2-ylmethoxy) stilbene ( 13c )

Compound 13c was prepared from 13b (260 mg, 0.7 mmol) using the same method as described for compound 15c . 13c (190 mg, 82%):

Example  45

4-nitro-4 '-(1-tosyl-3-hydroxy-propan-2-ylmethoxy) stilbene ( 13d )

Compound 13d was prepared from 13c (80 mg, 0.24 mmol) using the same method as described for compound 15d . 13d (66 mg, 56%):

Example  46

4-nitro-4 '-(1-fluoro-3-hydroxy-propan-2-ylmethoxy) stilbene ( 13e ).

Compound 13e was prepared from 13d (33 mg, 0.069 mmol) using the same method as described for compound 15e . 13e (20 mg, 88%):

Example 47

4-amino-4 '-(1-fluoro-3-hydroxy-propan-2-ylmethoxy) stilbene ( 14e ).

Compound 14e was prepared from 13e (37 mg, 0.11 mmol) using the same method as described for compound 14a . 14e (24 mg, 71%):

Example  48

4-N-methyl-amino-4 '-(1-fluoro-3-hydroxy-propan-2-ylmethoxy) stilbene ( 16e )

Under nitrogen atmosphere, sodium methoxide (22 mg, 0.4 mmol), then paraformaldehyde (12 mg, 0.4 mmol) was added to a suspension of compound 14e (24 mg, 0.08 mmol) in methanol (6 mL). The solution was heated to reflux for 2 hours and cooled to 0 ° C. using an ice bath. Sodium borohydride (15 mg, 0.4 mmol) was added in several portions. The reaction mixture was again refluxed for 1 hour and poured into crushed ice cubes. After standard workup with dichloromethane, the residue was subjected to TLC for silica gel preparation (4.5% methanol in dichloromethane) to give product 16e (23 mg, 92%).

Example  49

4-N-methyl-amino-4'-hydroxy stilbene ( 16a )

Compound 16a was prepared from 14a (105 mg, 0.5 mmol) using the same method as described for compound 16e . 16a (100 mg, 89%):

Example  50

12 (n = 6, 8) General microwave process for the production of stilbenes

Microwave synthesis: A mixture of 16a , alkylating agent (1 equiv) and K ₂ CO ₃ (3 equiv) in DMF (1 mL / 0.05 mmol SB-13) was placed in a sealed tube and heated in a microwave oven under the following conditions: 180 ° C., 10 minutes, high absorption level. The solvent was then removed and the desired product was obtained by PTLC [CH ₂ Cl ₂ -MeOH (97: 3) as developing solvent) (Yield: 42-60% depending on the alkylating agent used).

Example  51

(4- (2- (4- (2- (2- (2- (2- (2- (2-fluoro-ethoxy) -ethoxy) -ethoxy) -ethoxy) -ethoxy) -ethoxy)- Ethoxy) -phenyl) -vinyl) -phenyl) -methyl-amine ( 12 , n = 6):

Yield 60%.

Example  52

(4- (2- (4- (2- (2- (2- (2- (2- (2- (2- (2-fluoro-ethoxy) -ethoxy) -ethoxy) -ethoxy) -ethoxy) -ethoxy ) -Ethoxy) -ethoxy) -ethoxy) -ethoxy) -phenyl) -vinyl) -phenyl) -methyl-amine ( 12 , n = 8):

Yield 42%.

Example  53

Preparation of Brain Tissue Homogenates

Postmortem brain tissue was obtained from AD patients at necropsy and neuropathological diagnosis was confirmed by current criteria (NIA-Reagan Institute Consensus Group, 1997). Homogenates were then prepared from the cleaved gray matter of AD patients at a concentration of about 100 mg wet tissue / ml in phosphate buffered saline (PBS, pH 7.4) (at a setting of 6 for 30 seconds in a motor-operated glass homogenizer). . Homogenates were aliquoted in 1 ml aliquots and stored at −70 ° C. for 6-12 months without loss of binding signal.

Example 54

Combined research

As previously reported, [ ¹²⁵ I] IMPY having a specific activity of 2,200 Ci / mmol and greater than 95% radioactive compound purity was prepared using a standard iododestannylation reaction and simplified C Purification by -4 minicolumn (Kung MP, et al., Euro J Nucl Med Mol Imag. 2004; 31: 1136-45). Binding assays were performed in 12 x 75 mm borosilicate glass tubes. The reaction mixture is 50 μl (20-50 μg) brain homogenate, 50 μl [ ¹²⁵ I] IMPY (diluted at 0.04-0.06 nM in PBS) and 10 ^{−5 in} PBS containing 0.1% bovine serum albumin BSA. -10 ^-10 M diluted in search sequence) contained 50 ㎕ the final volume 1 ml. Nonspecific binding is defined as the presence of IMPY (600 nM) in the same assay. The mixture was incubated at 37 ° C. for 2 hours, and then the bound radioactive material and free radioactive material were passed through a Whatman GF / B filter using a Brandel M-24R cell harvester. After separation by vacuum filtration, it was washed twice with 3 ml of PBS at room temperature. Filters containing bound ¹²⁵ I ligands were assayed for radioactive content with a counting efficiency of 70% in a gamma counter (Packard 5000). Under assay conditions, specifically bound fractions were less than 15% of total radioactive material. For the inhibition test results, non-linear regression analysis was performed using the EBDA used to calculate K _i values. The results are shown in Table 1 below.

Each value was obtained from three independent measurements performed in duplicate.

Fluorinated PEG stilbenes ( 12a-d ) showed good binding affinity (K _i = 2.9-6.7 nM), and the corresponding hydroxyl substituent analogs ( 3a-d ) also showed very high binding affinity (K _i = 2.8-5.2 nM) (Table 1). The lipophilic properties of this series of labeled formulations [ ¹⁸ F] 12a-d were in the appropriate range (logP values were 2.52, 2.41, 2.05 and 2.28 for n = 2 to 5, respectively). PEG groups can control molecular size and distance between fluorine atoms and stilbene central structures without affecting Aβ plaque-specific binding affinity.

Example  55

Film Self-Recording Method

Brains were frozen in powder dry ice to obtain brain sections of AD patients and cut into 20 μm thick sections. Sections were incubated with [ ¹⁸ F] tracers (200,000-250,000 cpm / 200 μl) at room temperature for 1 hour. The sections were then immersed in saturated Li ₂ CO ₃ solution in 40% EtOH (2 washes twice), washed with 40% EtOH (1 wash 2 minutes) and then with water for 30 seconds. After drying, ¹⁸ F-labeled sections were exposed to Kodak MR films overnight.

Example  56

In vivo plaque labeling with [ ¹⁸ F] 12b and [ ¹⁸ F] 12d

In vivo evaluation was performed using either dual transgenic APP / PS1 or single transgenic APP2576 mice provided in favor of AstraZeneca. After anesthesia with 1% isoflurane, [ ¹⁸ F] 12b or [ ¹⁸ F] 12d 250-300 μCi in 200 μl of 0.1% BSA solution was injected through the tail vein. The animals were allowed to recover for 60 minutes and then sacrificed with the head. Brains were immediately isolated and frozen in powder dry ice. 20 μm sections were cut and exposed to Kodak MR film overnight. Thus, an ex vivo film self-radiation record was obtained.

Example 57

Long-term Distribution of Normal Mice

Under isoflurane anesthesia, 0.15 mL of 0.1% bovine serum albumin solution containing [ ¹⁸ F] tracer (5-10 μCi) was injected directly into the tail vein of ICR mice (22-25 g, male). 120 minutes after injection, mice (n = 3 for each time point) were sacrificed by cervical dislocation. Subject organs were isolated and weighed and then assayed for radioactivity on radioactivity content using an automatic gamma counter. Percent doses per organ were calculated by comparing the aliquots of the appropriately diluted injected material with the number of tissue counts. Total activity of blood was calculated assuming blood was 7% of total body weight. The percent dose / g of the sample was calculated by comparing the count of tissue counts with the count of diluted initial dose.

Radioactive compounds containing [ ¹⁸ F] 12a-d penetrated the intact blood-brain barrier, showing good brain uptake (6.6-8.1% dose / g brain) in normal mice 2 minutes after intravenous injection (Table 2A and B). Because the use of mice in the biodistribution experiments, these young mouse brain in was expected that Aβ plaque is not observed, therefore, the labeled agent ^[18 F] 12a-d were removed quickly from the brain 60 minutes injected intravenously (1.2-2.6% dose / g brain). High initial absorption and rapid clearance in normal mouse brains (not including Aβ plaques in the brain) are highly desirable properties for Aβ plaque-targeted imaging agents. The values reported in Table 2 are comparable to those reported for [ ¹¹ C] PIB and [ ¹¹ C] SB-13 (Mathis CA, et al., Curr Pharm Des. 2004; 10: 1469-92; Ono M , et al., Nucl Med Biol. 2003; Mathis CA, et al., J Med Chem. 2003).

The detailed biodistribution of [ ¹⁸ F] 12b is shown in Table 2A. Two minutes after injection, the compound is absorbed into the liver, kidneys, lungs and muscles, which appears to reflect a general blood perfusion pattern. Bone uptake was high at 120 minutes (2.74% dose / g), suggesting that there may be in vivo defluorination. However, free fluorine is not absorbed by brain tissue, so bone uptake is relatively low. The other PEG stilbene derivatives, 12a, c, d , showed similar biodistribution patterns (Table 2B).

Example 58

Partition coefficient

In vitro, partition coefficients were determined by mixing [ ¹⁸ F] tracers with 3 g each of 1-octanol and buffer (0.1 M phosphate, pH 7.4). The test tubes were vortexed at room temperature for 3 minutes and then centrifuged for 5 minutes. Two weighed samples (0.5 g each) from 1-octanol and buffer layers were counted in a well counter. The partition coefficient was determined by calculating the cpm / g of 1-octanol to cpm / g of buffer. Samples of the 1-octanol layer were redistributed (typically three or four distributions) until a uniform partition coefficient value was obtained. Measurements were carried out in triplicates and repeated three times.

Those skilled in the art will understand that the same can be accomplished within a broad and equivalent range of conditions, formulations and other parameters without affecting the scope of the invention or any of its embodiments. All patents, patent applications, and publications cited herein are incorporated by reference in their entirety.

Claims (37)

A compound of formula (I), a pharmaceutically acceptable salt or drug precursor thereof. <Formula I>

Where R ¹ is selected from the group consisting of a to k; a. NR ^a R ^b (wherein R ^a is and R ^b is independently hydrogen, C _{1 -4} alkyl, or (CH _{2) d} F ^18, d is an integer of 1 to 4), b. Hydroxy, c. C _{1 - 4} alkoxy, d. Hydroxy (C _1-4 ) alkyl, e. halogen, f. Cyano, g. Hydrogen, h. Nitro, i. (C ₁ -C ₄ ) alkyl, j. Halo (C ₁ -C ₄ ) alkyl, and k. Formyl; R ^{1 '} is selected from the group consisting of a to f; a. ¹²³ I, ¹²⁵ I, ¹³¹ I, ¹⁸ F or ⁷⁶ Br, b. Hydrogen, c. ¹⁸ F (C _{1 -4)} alkyl, d. ^[18 F (C _{1 -4)} alkyl] amino, e. [ ¹⁸ F (C ₁ -C ₄ ) alkyl] alkylamino, and f. ¹⁸ F (C ₁ -C ₄ ) alkoxy; R ² is selected from the group consisting of i to iv; i. Hydroxyl, C _{1 - 4} alkoxy, (C ₁ -C ₄₎ - alkyl-oxo-alk (C ₁ -C ₄₎ aryloxy, (C ₁ -C ₄₎ - alkyl, oxo (C ₁ -C ₄₎ - alkyl, oxo ( C ₁ -C ₄ ) alkoxy, (C ₁ -C ₄ ) -alkyloxo (C ₁ -C ₄ ) -alkyloxo (C ₁ -C ₄ ) -alkyloxo (C ₁ -C ₄ ) alkoxy, carboxy (C ₁ -C ₄₎ alkyl, halo (C ₁ -C ₄₎ alkoxy, halo (C ₁ -C ₄₎ - alkyl, oxo (C ₁ - C ₄₎ alkoxy, halo (C ₁ -C ₄₎ alkyl, oxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) -alkyloxy, halo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxy, halo (C ₁ -C ₄ ) alkyl, NR ⁶ R ^{6 ′} Wherein R ⁶ and R ^{6 ′} are independently selected from the group consisting of hydrogen, hydroxy (C ₁ -C ₄ ) alkyl and C ₁ -C ₄ alkyl), phenyl (C ₁ -C ₄ ) alkyl, ¹⁸ F ( C ₁ -C ₄ ) alkoxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkoxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C _1- C ₄ ) alkyloxy, ¹⁸ F (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxo (C ₁ -C ₄ ) alkyloxy or ¹⁸ F ( C ₁ -C ₄ ) alkyl; ii.

(Wherein, q is an integer from 1 to 10; Z is ¹⁸ F, ¹⁸ F-substituted benzoyloxy, ¹⁸ F-substituted (C _{1 -4)} alkoxy, ¹⁸ F-substituted benzyloxy, optionally ¹⁸ F- phenoxy hour, ¹⁸ F-substituted phenyl (C _1-4) alkyl, substituted aryloxy, and ¹⁸ F ¹⁸ F-substituted C _{6 -10} aryl, optionally is selected from the group consisting of ¹⁸ F- phenyl; R ^30, R ³¹ , and R ³² and R ³³ is selected from hydrogen, hydroxy, C _{1 -4} alkoxy, C _{1 -4} alkyl and hydroxy (C _1-4) alkyl group consisting of independently, in each case.) iii.

(Wherein Z, R ³⁰ , R ³¹ , R ³² and R ³³ are as described above). iv.

Wherein Y is ¹⁸ F, ¹⁸ F substituted benzoyloxy, ¹⁸ F substituted phenyl (C _1-4 ) alkyl, ¹⁸ F substituted aryloxy, optionally ¹⁸ F-phenoxy and ¹⁸ F substituted C _{6 -10} aryl, optionally ¹⁸ F-phenyl; U is hydrogen, hydroxy, ¹⁸ F, ¹⁸ F substituted benzoyloxy, ¹⁸ F substituted phenyl (C _1-4 ) alkyl, ¹⁸ F substituted aryloxy, optionally ¹⁸ F-phenoxy and ¹⁸ F substituted C _6-10 aryl, optionally is selected from the group consisting of ¹⁸ F- phenyl; ^{R 34, R 35, R 36} , R 37, R 38, R 39 and R ⁴⁰ are independently at each occurrence hydrogen, hydroxy, C _1-4 alkoxy, C _{1 -4} alkyl and hydroxy (C _1-4 ) Is selected from the group consisting of alkyl); R ⁷ and R ⁸ are independently at each occurrence a halogen, hydrogen, hydroxy, amino, methylamino, dimethylamino, C _{1 -4} alkoxy, C _{1 -4} alkyl and hydroxy (C _1-4) alkyl group consisting of And at least one of R ⁷ and R ⁸ is halogen. The method of claim 1, wherein R ^{1 'is} hydrogen, R ¹ is NR ^a and R ^b, R ^a and R ^b are independently hydrogen or C _{1 -4} alkyl. The compound of claim 2, wherein R ⁷ and R ⁸ are hydrogen or fluorine and at least one of R ⁷ and R ⁸ is fluorine. The compound of claim 2, wherein R ² is a group of the formula ii. ii.

Wherein Z, R ³⁰ , R ³¹ , R ³² and R ³³ are as described above. The compound of claim 4, wherein q is an integer from 2 to 5. 6. The compound of claim 5, wherein R ³⁰ , R ³¹ , R ³² and R ³³ are in each case hydrogen. The compound of claim 6, wherein q is an integer from 3 to 4. 8. 8. The compound of claim 7, wherein Z is ¹⁸ F. A compound according to claim 8 having the formula

Wherein one of R ⁷ and R ⁸ is hydrogen and the other is halogen. A compound according to claim 8 having the formula

Wherein one of R ⁷ and R ⁸ is hydrogen and the other is halogen. The compound of claim 2, wherein R ² is a group of formula iii. iii.

Wherein Z, R ³⁰ , R ³¹ , R ³² and R ³³ are as described above. 12. The compound of claim 11, wherein Z is ¹⁸ F. 13. Compounds according to claim 12, wherein R ³⁰ , R ³¹ , R ³² and R ³³ are in each case hydrogen. The compound of claim 2, wherein R ² is a group of formula iv. iv.

Wherein U is hydroxy. The compound of claim 14, wherein R ³⁴ , R ³⁵ , R ³⁶ , R ³⁷ , R ³⁸ , R ³⁹ and R ⁴⁰ are in each case hydrogen. The compound of claim 15 having the structure

Wherein one of R ⁷ and R ⁸ is hydrogen and the other is halogen. A compound of formula II <Formula II>

Where R ¹ is selected from the group consisting of a to d; a. NR ^a R ^b (where, R ^a and R ^b are independently hydrogen, C _{1 -4} alkyl, or (CH _{2) d} X (here standing, X is ¹⁸ F, or d is an integer of 1 to 4), R ^{both a} and R ^b are oxygen and form nitro), b. Hydroxy, c. C _{1 - 4} alkoxy, and d. Hydroxy (C _1-4 ) alkyl; R ² is selected from the group consisting of i to iii; i.

(Wherein, q is an integer of from 2 to 10; Z is ¹⁸ F, ¹⁸ F-substituted benzoyloxy, ¹⁸ F-substituted (C _{1 -4)} alkoxy, ¹⁸ F-substituted benzyloxy, substituted phenyl ¹⁸ F ( C _1-4) alkyl, ¹⁸ F and ¹⁸ F-substituted aryloxy-substituted C _{6 -10} is selected from the group consisting of ^{aryl; R 30, R 31, R} 32 and R ³³ are independently at each occurrence hydrogen, hydroxy hydroxy, C _{1 -4} alkoxy, C _{1 -4} alkyl and hydroxy _is selected from (C _{1 4)} alkyl group consisting of a). ii.

(Wherein Z, R ³⁰ , R ³¹ , R ³² and R ³³ are as described above). iii.

(Wherein, Y is ¹⁸ F, ¹⁸ F-substituted benzoyloxy, ¹⁸ F-substituted phenyl (C _1-4) alkyl, ¹⁸ F-substituted selected from aryloxy and ¹⁸ F-substituted C _{6 -10} aryl group consisting of Become; U is from the group consisting of hydrogen, hydroxy, ¹⁸ F, ¹⁸ F-substituted benzoyloxy, ¹⁸ F-substituted phenyl (C _1-4) alkyl, ¹⁸ F and ¹⁸ F-substituted aryloxy-substituted C _{6 -10} aryl group consisting of Selected; ^{R 34, R 35, R 36} , R 37, R 38, R 39 and R ⁴⁰ are independently at each occurrence hydrogen, ¹⁸ F, hydroxy, C _{1 -4} alkoxy, C _{1 -4} alkyl and hydroxy (C _1-4 ) alkyl.); R ⁷ and R ⁸ are independently selected from hydrogen, hydroxy, amino, methylamino, dimethylamino, C _{1 -4} alkoxy, C _{1 -4} alkyl and hydroxy (C _1-4) alkyl at each occurrence the group consisting of do. The compound of claim 17, wherein R ¹ is NR ^a R ^b and R ^a And R ^b is independently hydrogen or C _{1 -4} alkyl. 19. The compound of claim 18, wherein R ² is a group of formula i. i.

Wherein Z, R ³⁰ , R ³¹ , R ³² and R ³³ are as described above. 20. The compound of claim 19, wherein q is an integer from 2 to 5. The compound of claim 20, wherein R ⁷ and R ⁸ are each hydrogen. The compound of claim 21, wherein R ³⁰ , R ³¹ , R ³² and R ³³ are in each case hydrogen. The compound of claim 22 having the formula

The compound of claim 22 having the formula

The compound of claim 18, wherein R ² is a group of formula ii. ii.

Wherein Z, R ³⁰ , R ³¹ , R ³² and R ³³ are as described above. The compound of claim 25, wherein Z is ¹⁸ F. ²⁷ . The compound of claim 26, wherein R ⁷ and R ⁸ are each hydrogen. The compound of claim 27, wherein R ³⁰ , R ³¹ , R ³² and R ³³ are in each case hydrogen. The compound of claim 18, wherein R ² is a group of formula iii. iii.

Wherein U is hydroxy. ^30. The compound of claim 29, wherein R ³⁴ , R ³⁵ , R ³⁶ , R ³⁷ , R ³⁸ , R ³⁹ and R ⁴⁰ are in each case hydrogen. The compound of claim 30 having the structure

A compound of formula III <Formula III>

Wherein n is an integer from 1 to 4; R ⁷ and R ⁸ are independently selected from hydrogen, hydroxy, amino, methylamino, dimethylamino, C _{1 -4} alkoxy, C _{1 -4} alkyl and hydroxy (C _1-4) alkyl at each occurrence the group consisting of Become; R ⁴¹ is selected from the group consisting of hydroxy and NR ^a R ^b , wherein R ^a and R ^b are independently hydrogen or C 1-4 alkyl, or both R ^a and R ^b are oxygen and form nitro. 33. The compound of claim 32, wherein n is 1 and R ⁴¹ is selected from the group consisting of hydroxy, methylamino and dimethylamino. A pharmaceutical composition comprising the compound of any one of claims 1, 17 and 32. 33. A diagnostic composition for imaging an amyloid deposit, comprising the radiolabelled compound of any one of claims 1, 17 and 32. a. Introducing a detectable amount of the diagnostic composition of claim 35 into a mammal; b. Allowing sufficient time for the labeled compound to bind to the amyloid deposit; And c. Detecting the labeled compound in association with one or more amyloid deposits Imaging method of amyloid deposits comprising a. A method of inhibiting amyloid plaque aggregation in a mammal comprising administering an amount of the composition of claim 34 to inhibit amyloid plaque aggregation.

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